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α硫辛酸对高糖培养神经膜细胞氧化应激及凋亡的影响
The effects of Alpha lipoic acid on apoptosis and oxidative stress of Schwann cell induced by High glucose in vitro
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DOI:
作者:
秦利
QIN LI
作者单位:
上海交通大学医学院附属新华医院内分泌科
Department of Endocrinology,Xinhua Hospital,Shanghai Jiaotong University School of medicine
关键词:
糖尿病周围神经病变;神经膜细胞;氧化应激;α-硫辛酸
Diabetic peripheral neuropathy, Schwann cell, Oxidative stress , Alpha lipoic acid
摘要:
[摘要] 目的 探讨α硫辛酸对高糖培养神经膜细胞(SCs)氧化应激及凋亡的影响。方法 原代培养并纯化SD乳鼠坐骨神经SCs,S-100蛋白免疫组织化学法进行鉴定。取第3~4代SCs为研究对象,分为正常组、高糖组、波动高糖组及不同浓度的α硫辛酸组,干预48h后MTT法检测细胞活力、TUNEL法检测细胞凋亡,Western blot检测Bax蛋白的表达,同时检测各组SCs的丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性。结果 (1) 稳定高糖及波动高糖均提高了SCs的氧化应激水平,升高凋亡相关Bax蛋白的表达,增加了细胞凋亡(P<0.05);(2)波动高糖较稳定高糖的上述效应更为明显(P<0.05);(3): α硫辛酸可以剂量依赖性改善波动高糖及稳定高糖对SCs的氧化损伤,抑制凋亡相关Bax蛋白的表达 (P<0.05)。结论 α-硫辛酸通过抑制氧化应激而改善高糖导致的SCs凋亡,该机制可能在改善糖尿病周围神经病变中起重要作用。
[Abstract] Objective To investigate the effects of Alpha lipoic acid (ALA) on apoptosis and oxidative stress of Schwann cells(SCs) cultured with high glucose medium in vitro. Methods SCs were primarily-cultured with high glucose medium in vitro. Methods SCs were primarily-cultured and purified from the sciatic nerves of newborn SD rats. S-100 protein immunohistochemical method was adopted for the identification of them. SCs of the 3rd~4th passage were collected and cultured with different mediums. They were divided into a normal group,a high glucose group,a intermittent high glucose group and intervention group in the presence of various concentrations of ALA. After 48 hours,MTT was adopted to detect the ability of SCs. Apoptosis was confirmed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method. Western blot were performed to analyze the expression levels of bax protein. In addition, the oxidative stress indexes in SCs, such as the content of MDA and the activity of SOD were also measured. Results (1) Compared with that of the normal glucose exposure, HG treatment significantly increased the oxidative stress level, up-regulated the expression of Bax protein, and increased the percentages of apoptotic cells in SCs.(2) The effect of IHG was significantly more potent than that of HG.(3) Treatment with ALA inhibited the HG and IHG-induced oxidative stress and down-regulation of Bax protein expression, inhibited the activation of molecular pathways of apoptosis in SCs in a dose-dependent manner. Conclusion ALA can antagonized the IHG and HG-induced apoptosis in SCs through inhibiting oxidative stress, which may play a key role in improving diabetic peripheral neuropathy.
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